Antibody was affinity purified using an epitope specific to MLL1 immobilized on solid support. The epitope recognized by A300-374A maps to a region between residues 2725 and 2775 of human myeloid/lymphoid or mixed-lineage leukemia 1 using the number given in Swiss-Prot entry Q03164 (GeneID 4297). The epitope is found in the C-terminal 180 kDa fragment generated by proteolytic cleavage. The epitope is found in isoform 14P-18B of MLL1. Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
ALL-1, CXXC7, CXXC-type zinc finger protein 7, histone-lysine N-methyltransferase 2A, HRX, HTRX1, lysine (K)-specific methyltransferase 2A, lysine N-methyltransferase 2A, mixed lineage leukemia 1, MLL, MLL1, MLL1A, Myeloid/lymphoid or mixed-lineage leukem
between 2725 and 2775
2 - 8°C
Detection of human MLL1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, and Jurkat cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-MLL1 antibody A300-374A (lot A300-374A-5) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
Detection of human MLL1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HEK293T cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-MLL1 antibody A300-374A (lot A300-374A-5) used for IP at 3 µg per reaction. MLL1 was also immunoprecipitated by rabbit anti-MLL1 antibody BL1412. For blotting immunoprecipitated MLL1, A300-374A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
ChIP-chip scatter plot of anti-MLL1 (A300-374A) enriched DNA binding sites versus input reference DNA. A. 10 µg of A300-374A was used to immunoprecipitate chromatin from K-562 cells according to Ren et al (Genes Dev. 2002 16: 245-256). immunoprecipitatesd DNA and reference DNA were amplified via ligation-mediated PCR and the products labeled with fluorescent dUTPs. The labeled ChIP and reference DNA were pooled, hybridized to a DNA microarray, and analyzed. Data points below the +3 SD curve (red line) represent significantly enriched binding sites. B. As a control, a similar experiment was performed using normal rabbit IgG. Compared to the anti-MLL1 ChIP, normal rabbit IgG showed little enrichment.